3D Cell Culture

Preface...
Table of Contents...
Contributing Authors...

1.3D Cell Culture: Techniques For and Beyond Organoid Applications 
Zuzana Sumbalova Koledova

Part I Hydrogels and Scaffolds for 3D Cell Culture

2.Preparation of Corneal Tissue Matrix Bioink for 3D Bioprinting 
Shibu Chameettachal and Falguni Pati   

3.Decellularization of Pig Lung to Yield Three-Dimensional Scaffold for Lung Tissue Engineering 
Katarína Čimborová, Hana Kotasová, Vendula Pelková, Veronika Sedláková, and Aleš Hampl

4.Three-Dimensional Hanging Drop Spheroid Plates for Easy Chimeric Antigen Receptor (CAR) T Cytotoxicity Assay
Zhenzhong Chen and Sungsu Park 

5.Static 3D Osteoblast Cell Culture on 3D Printed Titanium Scaffolds 
Carolina Oliver-Urrutia, Anna Diez-Escudero, Zuzana Sumbalova Koledova, Ladislav Čelko, Nils P. Hailer, and Edgar B. Montufar

6.Modelling Acute Myeloid Leukaemia using StarPEG-Heparin Hydrogels 
P. Lewen Holloway, Akhilandeshwari Ravichandran, Julien Clegg, Claudia Bruedigam, and Laura J. Bray

Part II 3D Organoid and Organotypic Cultures

7.A Decision Tree to Guide Human and Mouse Mammary Organoid Model Selection
Marika Caruso, Kamyab Saberiseyedabad, Larissa Mourao, and Colinda L.G.J. Scheele

8.Fibroblast-Epithelium Co-Culture Methods using Epithelial Organoids and Cell Line-Derived Spheroids 
Jakub Sumbal and Zuzana Sumbalova Koledova

9.Differentiation of Fibroblasts to Adipocytes in 3D for a Co-Culture with Mammary Organoids and Immunohistological Analysis 
Matea Brezak, Lukas Kubec, and Zuzana Sumbalova Koledova

10.Organotypic 3D Cell Culture of the Embryonic Lacrimal Gland  
Alison Kuony, Matea Brezak, René-Marc Mège, and Zuzana Sumbalova Koledova 

11.Renal Organoids from Whole Kidney Cells 
Liang Chen

12.Organotypic 3D Ex Vivo Co-Culture Model of the Macro-Metastasis/Organ Parenchyma Interface
Rachel Blazquez, Daniela Sparrer, Jessica Sonbol, Jürgen Philipp, Florian Schmieder, and Tobias Pukrop 

Part III Functional Experiments in 3D Cultures

13.Addressing the Mechanical Interaction between Cancer-Associated Fibroblasts and Cancer Cells by Laser Ablation 
Carlos Perez-Gonzalez and Jorge Barbazán 

14.Double-Barrel Perfusion System for Modification of Luminal Contents of Intestinal Organoids 
Nicholas J. Ginga and Raleigh Slyman 

15.Mechanical Actuation of Organoids in Synthetic Microenvironments 
Francesca Sgualdino, Lorenzo Mattolini, Brian Daza Jimenez, Kieran Patrick, Abdel Rahman Abdel Fattah, and Adrian Ranga 

Part IV Microfluidic and Bioprinting Approaches for 3D Cell Culture

16.3D Breast Cancer Model on Silk Fibroin-Integrated Microfluidic Chips 
Eylul Gulsen Yilmaz and Fatih Inci

17.Tumor-Microenvironment-on-Chip Platform for Assessing Drug Response in 3D Dynamic Culture
Hakan Berk Aydin, Hye-ran Moon, Bumsoo Han, Altug Ozcelikkale, and Ahmet Acar 

18.Suspended 3D Bioprinting for Engineering Miniaturized In Vitro Tumor Models 
Maria V. Monteiro, Marta Rocha, Vítor M. Gaspar, and João F. Mano 

Part V Imaging and Image Analysis of 3D Cell Cultures

19.Real-Time Cell Cycle Imaging in a 3D Cell Culture Model of Melanoma, Quantitative Analysis, Optical Clearing, and Mathematical Modeling
Loredana Spoerri, Kimberley A. Beaumont, Andrea Anfosso, Ryan J. Murphy, Alexander P. Browning, Gency Gunasingh, and Nikolas K. Haass

20.Quantitative Analysis of Whole-Mount Fluorescence-Stained Tumor Spheroids in Phenotypic Drug Screens 
Elina Nuernberg, Roman Bruch, Mathias Hafner, Ruediger Rudolf, and Mario Vitacolonna

Subject Index List...

Since the publication of the previous edition, the use of 3D cell and organoid cultures has become more widespread across laboratories. This second edition volume expands on the previous edition with discussions about the latest organoid models developed for many more organs; new hydrogels and devices for 3D culture; and the organoid systems that have been improved by incorporating more components of tissue microenvironments in the in vitro culture. The chapters in this book are organized into five parts and cover topics such as biofabrication, organoids, microfluidic systems, bioprinting, and image analysis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.